EP4332213A1 - Wachstumsförderer für milchsäurekokken - Google Patents

Wachstumsförderer für milchsäurekokken Download PDF

Info

Publication number
EP4332213A1
EP4332213A1 EP22795395.7A EP22795395A EP4332213A1 EP 4332213 A1 EP4332213 A1 EP 4332213A1 EP 22795395 A EP22795395 A EP 22795395A EP 4332213 A1 EP4332213 A1 EP 4332213A1
Authority
EP
European Patent Office
Prior art keywords
lactic acid
medium
acid
coccus
dicarboxylic acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP22795395.7A
Other languages
English (en)
French (fr)
Inventor
Yu HIROSE
Hazuki MURAKAWA
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kirin Holdings Co Ltd
Original Assignee
Kirin Holdings Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from JP2021101500A external-priority patent/JP7248250B2/ja
Application filed by Kirin Holdings Co Ltd filed Critical Kirin Holdings Co Ltd
Publication of EP4332213A1 publication Critical patent/EP4332213A1/de
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/46—Streptococcus ; Enterococcus; Lactococcus

Definitions

  • the present invention relates to a growth promoter for a lactic acid coccus.
  • the present invention also relates to a medium for a lactic acid coccus, a method for cultivation of a lactic acid coccus, and a method for promoting the growth of a lactic acid coccus.
  • LB medium and MRS medium are generally used in the cultivation of lactic acid cocci.
  • sufficient growth of lactic acid cocci may not be achieved, depending on the type of medium.
  • the methods described in Patent Documents 1 and 2 have been known so far as methods for efficient cultivation of a lactic acid coccus.
  • the present inventors have found that, in the cultivation of a lactic acid coccus, the growth of the lactic acid coccus is promoted by including a dicarboxylic acid or a salt thereof in a medium.
  • the present invention is based on this finding.
  • the growth of a lactic acid coccus can be promoted by including a dicarboxylic acid or a salt thereof in a medium to cultivate the lactic acid coccus, and, therefore, the present invention is advantageous in that the lactic acid coccus can be cultivated easily and efficiently.
  • the present invention is advantageous also in that the growth of a lactic acid coccus can be promoted by including a dicarboxylic acid or a salt thereof also in a medium that is free of meat extract.
  • the growth promoter of the present invention can be used in the cultivation of a lactic acid coccus.
  • the lactic acid coccus as the target for growth promotion is not particularly limited, and examples thereof include lactic acid cocci belonging to the genus Lactococcus, the genus Leuconostoc, the genus Pediococcus, and the genus Streptococcus.
  • Lactococcus lactis subsp. lactis examples include Lactococcus lactis subsp. lactis, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. hordniae, and Lactococcus lactis subsp. lactis is preferable.
  • bacteria belonging to the genus Lactococcus include Lactococcus lactis subsp. lactis JCM5805, Lactococcus lactis subsp. lactis NBRC12007, Lactococcus lactis subsp. lactis NRIC1150, Lactococcus lactis subsp. lactis JCM20101, Lactococcus lactis subsp. lactis JCM7638, Lactococcus lactis subsp. lactis ATCC11454, Lactococcus garvieae NBRC100934, Lactococcus lactis subsp. cremoris JCM16167, Lactococcus lactis subsp.
  • cremoris NBRC100676, Lactococcus lactis subsp. hordniae JCM1180, and Lactococcus lactis subsp. hordniae JCM11040, and Lactococcus lactis subsp. lactis JCM5805 is preferable.
  • bacteria belonging to the genus Leuconostoc include Leuconostoc lactis.
  • Specific examples of bacteria belonging to the genus Leuconostoc include Leuconostoc lactis NBRC12455.
  • bacteria belonging to the genus Pediococcus include Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus cellicola, Pediococcus claussenii, Pediococcus damnosus, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, and Pediococcus stilesii.
  • Specific examples of bacteria belonging to the genus Pediococcus include Pediococcus acidilactici JCM8797 and Pediococcus damnosus JCM5886.
  • Examples of bacteria belonging to the genus Streptococcus include Streptococcus thermophilus.
  • the JCM bacterial strains are available from Microbe Division, RIKEN BioResource Research Center (3-1-1 Koyadai, Tsukuba, Ibaraki); the NBRC bacterial strains are available from Biological Resource Center, National Institute of Technology and Evaluation (2-5-8 Kazusa-Kamatari, Kisarazu, Chiba); the NRIC bacterial strains are available from NODAI Culture Collection Center, Tokyo University of Agriculture (1-1-1 Sakuragaoka, Setagaya, Tokyo); and the ATCC bacterial strains are available from American Type Culture Collection (U.S.A.).
  • the Lactococcus lactis subsp. lactis JCM5805 strain is available from Microbe Division, RIKEN BioResource Research Center as described above, but, in the present invention, the same bacterial strain as the JCM5805 strain, that is stored at a culture collection other than Microbe Division, RIKEN BioResource Research Center, can be used. Specifically, the same bacterial strain as the JCM5805 strain is available from Biological Resource Center, National Institute of Technology and Evaluation (2-5-8 Kazusa-Kamatari, Kisarazu, Chiba), NODAI Culture Collection Center, Tokyo University of Agriculture (1-1-1 Sakuragaoka, Setagaya, Tokyo), American Type Culture Collection (U.S.A.), and the like.
  • the dicarboxylic acid which is the active ingredient of the growth promoter of the present invention, is not particularly limited as long as it is a compound having two carboxyl groups in its structure.
  • An aliphatic dicarboxylic acid can be preferably used.
  • the hydrocarbon chain that constitutes the aliphatic dicarboxylic acid is preferably a saturated hydrocarbon chain.
  • the aliphatic dicarboxylic acid is preferably an aliphatic dicarboxylic acid having 4 to 6 carbon atoms, more preferably an aliphatic dicarboxylic acid having 4 or 5 carbon atoms.
  • the aliphatic dicarboxylic acid having 4 to 6 carbon atoms includes, for example, a ketoglutaric acid ( ⁇ -ketoglutaric acid (sometimes referred to as " ⁇ KG” herein), ⁇ -ketoglutaric acid (acetone dicarboxylic acid)), malic acid, adipic acid, oxalic acid, succinic acid, maleic acid, and glutaric acid, and is preferably either or both of ⁇ -ketoglutaric acid and malic acid.
  • the dicarboxylic acid may be in the form of a salt
  • the salt include metal salts and ammonium salts.
  • the metal salts include alkali metal salts such as sodium salts and potassium salts.
  • the ammonium salts include salts of ammonium, tetramethylammonium and the like.
  • the salt of dicarboxylic acid include ⁇ -ketoglutaric acid sodium salt and malic acid sodium salt.
  • the growth promoter of the present invention can contain either or both of a dicarboxylic acid and its salt as the active ingredient.
  • the lower limit value (or more or more than) of the concentration (solid content concentration) of the dicarboxylic acid and/or salt thereof in a medium can be set to 0.01% by mass, 0.1% by mass, 0.25% by mass, or 0.5% by mass, and the upper limit value (or less or less than) thereof can be set to 8% by mass, 4% by mass, or 2% by mass. These lower and upper limit values can be combined arbitrarily.
  • the range of the concentration of the dicarboxylic acid and/or salt thereof in the medium can be set to, for example, 0.01 to 8% by mass, 0.1 to 4% by mass, 0.25 to 4% by mass, 0.5 to 4% by mass, 0.5 to 3% by mass, or 0.5 to 2% by mass.
  • the growth promoter of the present invention can promote the growth of a lactic acid coccus and efficiently cultivate the lactic acid coccus by setting the concentration of the dicarboxylic acid in the medium to fall within the concentration range defined above.
  • the growth promoter of the present invention can be added to a medium for cultivation of a lactic acid coccus for use.
  • the medium that can be used in the present invention is not particularly limited as long as it is a medium used in the cultivation of a lactic acid coccus, and examples thereof include MRS medium (deMan, Rogosa & Sharpe medium), M17 medium, and LB medium.
  • MRS medium deMan, Rogosa & Sharpe medium
  • M17 medium M17 medium
  • LB medium LB medium.
  • the growth promoter of the present invention may be added to the medium during cultivation of a lactic acid coccus, or may be added in advance before cultivation of a lactic acid coccus.
  • the medium that can be used in the present invention can contain meat extract as a supplementary nutritional ingredient. Examples of the meat extract include beef extract, pork extract, and chicken extract.
  • the medium that can be used in the present invention is preferably substantially free of meat extract, more preferably free of meat extract, from the viewpoint of environmental load, cost, food safety, and the like.
  • the form of the meat extract to be used include, for example, extract liquid, powder, and granule.
  • the content of the meat extract in the medium is calculated by dividing the solid content mass, which is a pure extract content, of the meat extract, by the mass of the medium and multiplying the obtained value by 100.
  • the medium which is free of meat extract can be appropriately prepared without using meat extract in the raw material for the medium exemplified above.
  • the growth promoter of the present invention can be added to the medium which is free of meat extract for use.
  • the conditions for cultivation of a lactic acid coccus using a medium added with the growth promoter of the present invention are not particularly limited, and normal conditions for cultivation of a lactic acid coccus can be employed.
  • the period for cultivation of a lactic acid coccus can be set to 20 to 48 hours, and is preferably 20 to 30 hours from the viewpoint of a growth phase.
  • the lower limit value (or higher or higher than) of the temperature for cultivation of a lactic acid coccus can be set to 20°C, 25°C, or 28°C, and the upper limit value (or lower or lower than) thereof can be set to 40°C, 35°C, or 30°C.
  • the cultivation temperature can be set to, for example, a range of 20 to 40°C, 20 to 35°C, 25 to 35°C, or 30 to 35°C, most preferably about 30°C.
  • the cultivation of a lactic acid coccus may be either liquid cultivation or solid cultivation.
  • the liquid cultivation can be performed using a fermenter whose conditions can be controlled, and examples thereof include stirring cultivation, aeration cultivation, shaking cultivation, and stationary cultivation.
  • the cultivation may be performed under either anaerobic conditions or aerobic conditions.
  • the stirring speed and aeration rate are appropriately selected according to the type of microbial cells and the cultivation conditions.
  • the pH of the medium that is used in the present invention may be any pH that does not affect the growth of a lactic acid coccus, and can be set to, for example, a range of 6.0 to 6.7.
  • the pH of the medium can be adjusted appropriately, and, for example, can be adjusted by adding a base such as sodium hydroxide or an acid such as phosphoric acid to the medium.
  • a culture of lactic acid coccus can be obtained by cultivating a lactic acid coccus in a medium added with the growth promoter of the present invention.
  • the obtained lactic acid coccus culture can be blended in foods and drinks as it is or after treatments such as concentration, dilution, drying, isolation and purification. That is, the growth promoter of the present invention can be used to promote the growth of a lactic acid coccus for the purpose of application to foods and drinks, and, from this viewpoint, a dicarboxylic acid and/or a salt thereof acceptable as a food can be used as the active ingredient in the present invention.
  • a medium for a lactic acid coccus containing a dicarboxylic acid and/or a salt thereof.
  • the medium for a lactic acid coccus of the present invention can be implemented according to the descriptions regarding the growth promoter of the present invention.
  • the medium for a lactic acid coccus of the present invention is advantageous in that it promotes the growth of a lactic acid coccus, and thus enables efficient cultivation of the lactic acid coccus.
  • a method for cultivation of a lactic acid coccus comprising the step of cultivation of a lactic acid coccus in the medium of the present invention.
  • the cultivation method of the present invention can be carried out according to the descriptions regarding the growth promoter of the present invention and the medium of the present invention.
  • a method for promoting the growth of a lactic acid coccus comprising the step of adding a dicarboxylic acid and/or a salt thereof to a medium for a lactic acid coccus.
  • the growth promoting method of the present invention can be carried out according to the descriptions regarding the growth promoter of the present invention.
  • a dicarboxylic acid and/or a salt thereof as a growth promoter for a lactic acid coccus, for promoting the growth of a lactic acid coccus, or in the method for promoting the growth of a lactic acid coccus of the present invention.
  • the use of the present invention can be carried out according to the descriptions regarding the growth promoter and growth promoting method of the present invention.
  • Example 1 the growth promoting effect of dicarboxylic acids on a lactic acid coccus was tested.
  • Lactococcus lactis subsp. lactis JCM5805 strain was used as the lactic acid coccus.
  • the medium used was MRS medium (MRS BROTH, CODE: CM0359, Oxoid; the composition thereof is indicated in Table 1; the same applies in Examples 3 to 6 of the present specification).
  • MRS medium MRS BROTH, CODE: CM0359, Oxoid; the composition thereof is indicated in Table 1; the same applies in Examples 3 to 6 of the present specification.
  • dicarboxylic acids ⁇ -ketoglutaric acid (FUJIFILM Wako Pure Chemical Corporation), disodium ⁇ -ketoglutarate (sometimes referred to as " ⁇ KG2Na” herein) (FUJIFILM Wako Pure Chemical Corporation) and malic acid (Tokyo Chemical Industry Co., Ltd.) were each added to the medium at a concentration (0.01 to 4.0% by mass) as shown in FIG.
  • Table 1 Composition of MRS medium g/L Peptone 10.0 Lab-Lemco powder (beef extract) 8.0 Yeast extract 4.0 Glucose 20.0 Sorbitan oleate 1 ml Dipotassium hydrogen phosphate 2.0 Sodium acetate trihydrate 5.0 Triammonium citrate 2.0 Magnesium sulfate heptahydrate 0.2 Manganese sulfate tetrahydrate 0.05 Purified water Balance
  • the medium used was MRS medium.
  • dicarboxylic acids ⁇ -ketoglutaric acid, disodium ⁇ -ketoglutarate, and malic acid were each added to the medium at a concentration 0.5 to 2.0% by mass) as shown in FIG. 2 , and sodium hydroxide was added so as to attain the same pH as that of a control (dicarboxylic acid-free medium) (pH: 6.2 ⁇ 0.2).
  • the bacterial cells were inoculated (seeded) so that the concentration of the bacterial cells was 0.1% (v/v) with respect to 10 ml of the medium, and the medium inoculated with the bacterial cells was placed in an automatic cultivation searching device OT-201 (Oriental Instruments; sometimes referred to as “bioplotter” herein), and cultivated at 30°C for 24 hours (stirring intensity condition: Medium).
  • OT-201 Oriental Instruments; sometimes referred to as "bioplotter” herein
  • the bacteria were counted by measuring the OD600 (turbidity) of the medium using an absorbance meter (Biochrom GeneQuant 1300).
  • FIGS. 1 and 2 The results were as shown in FIGS. 1 and 2 . It was confirmed that, in the anaerobic cultivation, the growth of the lactic acid coccus was significantly promoted in a concentration-dependent manner in the media added with ⁇ KG ( FIG. 1A ) and malic acid ( FIG. 1C ) as compared with the control (dicarboxylic acid-free medium). In addition, it was confirmed that the growth of the lactic acid coccus was significantly promoted in a concentration-dependent manner at a concentration of 0.1 to 4% by mass in the medium added with ⁇ KG2Na ( FIG. 1B ) as compared with the control (dicarboxylic acid-free medium). Also at 0.01% by mass, the growth of the lactic acid coccus tended to be promoted.
  • Example 2 the growth promoting effect of dicarboxylic acids on a lactic acid coccus was tested using a medium different from the medium used in Example 1.
  • Example 1 The lactic acid coccus described in Example 1, item (1), A was used.
  • the medium used was a medium obtained by adding 20 g/L of glucose to M17 medium (Difco M17 Broth, BD; the composition thereof is indicated in Table 2; the same applies hereinafter in the present specification) (sometimes referred to as "GM17 medium” herein).
  • M17 medium a medium obtained by adding 20 g/L of glucose to M17 medium
  • ⁇ -ketoglutaric acid and malic acid were each added to the medium at a concentration (0.5 to 2% by mass) as shown in FIG. 3 , and sodium hydroxide was added so as to attain the same pH as that of a control (dicarboxylic acid-free medium) (pH: 6.5 ⁇ 0.2).
  • the bacterial cells were inoculated (seeded) in a 15-ml conical tube (FALCON) so that the concentration of the bacterial cells was 0.1% (v/v) with respect to 10 ml of the prepared medium, and cultivated at 30°C for 24 hours in an incubator (TOKYO RIKAKIKAI CO., LTD.) (static cultivation).
  • Table 2 Composition of M17 medium Per 950 mL of purified water Pancreatic digest of casein 5.0 g Soy peptone 5.0 g Beef extract 5.0 g Yeast extract 2.5 g Ascorbic acid 0.5 g Magnesium sulfate 0.25 g Disodium ⁇ -glycerophosphate 19.0 g
  • the medium used was GM17 medium.
  • dicarboxylic acids ⁇ -ketoglutaric acid and malic acid were each added to the medium at a concentration (0.5 to 2% by mass) as shown in FIG. 4 , and sodium hydroxide was added so as to attain the same pH as that of a control (dicarboxylic acid-free medium) (pH: 6.5 ⁇ 0.2).
  • the bacterial cells were inoculated (seeded) so that the concentration of the bacterial cells was 0.1% (v/v) with respect to 10 ml of the prepared medium, and the medium inoculated with the bacterial cells was placed in an automatic cultivation searching device OT-201 (Oriental Instruments; sometimes referred to as “bioplotter” herein), and cultivated at 30°C for 24 hours (stirring intensity condition: Medium).
  • OT-201 Oriental Instruments; sometimes referred to as "bioplotter” herein
  • Example 3 the growth promoting effect of ⁇ -ketoglutaric acid on various lactic acid cocci was tested.
  • the lactic acid cocci indicated in Table 3 were used as the bacterial cells.
  • Table 3 List of lactic acid cocci Name of bacterial strain Bacterial species NBRC100934 Lactococcus garvieae JCM16167 Lactococcus lactis subsp. cremoris NBRC100676 Lactococcus lactis subsp. cremoris JCM1180 Lactococcus lactis subsp. hordniae JCM11040 Lactococcus lactis subsp. hordniae NBRC12007 Lactococcus lactis subsp. lactis NRIC1150 Lactococcus lactis subsp. lactis JCM20101 Lactococcus lactis subsp. lactis NBRC12455 Leuconostoc lactis JCM7638 Lactococcus lactis subsp. lactis ATCC11454 Lactococcus lactis subsp. lactis
  • Example 4 the growth promoting effect of ⁇ -ketoglutaric acid on lactic acid-producing bacilli belonging to the genus Lactobacillus was tested.
  • the lactic acid-producing bacilli of the genus Lactobacillus indicated in Table 4 were used as the bacterial cells.
  • Table 4 List of lactic acid-producing bacilli Name of bacterial strain Bacterial species LGG Lactobacillus rhamnosus GG JCM1112 Lactobacillus reuteri
  • Example 5 the growth promoting effect of a tricarboxylic acid on a lactic acid coccus was tested.
  • Example 1 Cultivation was performed in a similar manner as described in Example 1, item (1), B (i), except that citric acid (FUJIFILM Wako Pure Chemical Corporation) was added as a tricarboxylic acid, in place of the dicarboxylic acid, to the medium at a concentration (0.25 to 1% by mass) as shown in FIG. 7 (anaerobic cultivation).
  • citric acid FFUJIFILM Wako Pure Chemical Corporation
  • Example 6 the growth promoting effect of dicarboxylic acids on lactic acid cocci belonging to the genus Pediococcus was tested.
  • the lactic acid cocci of the genus Pediococcus indicated in Table 5 were used as the bacterial cells.
  • Table 5 List of lactic acid cocci Name of bacterial strain Bacterial species JCM8797 Pediococcus acidilactici JCM5886 Pediococcus damnosus
  • Example 1 Cultivation was performed in a similar manner as described in Example 1, item (1), B (i), except that ⁇ -ketoglutaric acid (concentrations shown in FIG. 8A (0.1 to 4% by mass) and FIG. 8C (1 to 4% by mass)) and malic acid (concentrations shown in FIG. 8B (0.1 to 2% by mass) and FIG. 8D (1 to 2% by mass)) were each added, as the dicarboxylic acids, to the medium (anaerobic cultivation).
  • ⁇ -ketoglutaric acid concentration shown in FIG. 8A (0.1 to 4% by mass) and FIG. 8C (1 to 4% by mass)
  • malic acid concentration shown in FIG. 8B (0.1 to 2% by mass) and FIG. 8D (1 to 2% by mass)
  • FIG. 8 The results were as shown in FIG. 8 . It was confirmed that the growth of the lactic acid cocci belonging to the genus Pediococcus was significantly promoted in a concentration-dependent manner in the media added with ⁇ KG ( FIGS. 8A and 8C ) and malic acid FIGS. 8B and 8D as compared with the control (dicarboxylic acid-free medium).
  • Example 7 the growth promoting effect of a dicarboxylic acid on lactic acid cocci in a meat extract-free medium was tested.
  • the lactic acid cocci indicated in Table 6 were used as the bacterial cells.
  • Table 6 List of lactic acid cocci Name of bacterial strain Bacterial species JCM5805 Lactococcus lactis subsp. lactis NRBC100934 Lactococcus garvieae JCM16167 Lactococcus lactis subsp. cremoris NBRC100676 Lactococcus lactis subsp. cremoris JCM1180 Lactococcus lactis subsp. hordniae JCM11040 Lactococcus lactis subsp. hordniae NBRC12007 Lactococcus lactis subsp. lactis NRIC1150 Lactococcus lactis subsp.
  • lactis JCM20101 Lactococcus lactis subsp. lactis NBRC12455 Leuconostoc lactis JCM7638 Lactococcus lactis subsp. lactis ATCC11454 Lactococcus lactis subsp. lactis
  • the medium used was MRS medium adjusted so as to have a composition as indicated in Table 7 (adjusted to have a pH of 6.2 by adding a phosphoric acid).
  • a phosphoric acid As the dicarboxylic acid, ⁇ -ketoglutaric acid was added to the medium at a concentration of 2.0% by mass, and sodium hydroxide was added so as to attain the same pH as that of a control (dicarboxylic acid-free medium) (pH: 6.2 ⁇ 0.2).
  • the bacterial cells were inoculated (seeded) in a 15-ml conical tube (FALCON) so that the concentration of the bacterial cells was 0.1% (v/v) with respect to 10 ml of the prepared medium, and cultivated at 30°C for 24 hours in an incubator (TOKYO RIKAKIKAI CO., LTD.) (static cultivation).
  • Table 7 Composition of MRS medium (free of meat extract) g/L Peptone (BD) 10.0 Yeast extract (BD) 4.0 Glucose (FUJIFILM Wako Pure Chemical Corporation) 20.0 Sorbitan oleate (Kanto Chemical Industry Co., Ltd.) 1 ml Dipotassium hydrogen phosphate (FUJIFILM Wako Pure Chemical Corporation) 2.0 Sodium acetate trihydrate (FUJIFILM Wako Pure Chemical Corporation) 5.0 Triammonium citrate (Kanto Chemical Industry Co., Ltd.) 2.0 Magnesium sulfate heptahydrate (FUJIFILM Wako Pure Chemical Corporation) 0.2 Manganese sulfate pentahydrate (Kishida Chemical Co., 0.05 Ltd.) Purified water Balance
  • Example 8 Evaluation of influence of meat extract in medium on growth of lactic acid coccus
  • Example 8 the influence of meat extract in a medium on the growth of lactic acid cocci was tested.
  • the lactic acid cocci indicated in Table 8 were used as the bacterial cells.
  • Table 8 List of lactic acid cocci Name of bacterial strain Bacterial species JCM5805 Lactococcus lactis subsp. lactis NRBC 100934 Lactococcus garvieae JCM16167 Lactococcus lactis subsp. cremoris NBRC 12007 Lactococcus lactis subsp. lactis NBRC12455 Leuconostoc lactis JCM7638 Lactococcus lactis subsp. lactis ATCC11454 Lactococcus lactis subsp. lactis
  • the meat extract-free medium used was MRS medium adjusted so as to have a composition as indicated in Table 7 (adjusted to have a pH of 6.2 by adding a phosphoric acid because the initial pH was around 7.0).
  • the meat extract-containing medium used was MRS medium adjusted so as to have a composition as indicated in Table 9 (adjusted to have a pH of 6.2 by adding a phosphoric acid).
  • the bacterial cells were inoculated (seeded) in a 15-ml conical tube (FALCON) so that the concentration of the bacterial cells was 0.1% (v/v) with respect to 10 ml of the prepared medium, and cultivated at 30°C for 24 hours in an incubator (TOKYO RIKAKIKAI CO., LTD.) (static cultivation).
  • Table 9 Composition of MRS medium (containing meat extract) g/L Peptone (BD) 10.0 Lab-Lemco powder (beef extract) (Oxoid) 8.0 Yeast extract (BD) 4.0 Glucose (FUJIFILM Wako Pure Chemical Corporation) 20.0 Sorbitan oleate (Kanto Chemical Industry Co., Ltd.) 1 ml Dipotassium hydrogen phosphate (FUJIFILM Wako Pure Chemical Corporation) 2.0 Sodium acetate trihydrate (FUJIFILM Wako Pure Chemical Corporation) 5.0 Triammonium citrate (Kanto Chemical Industry Co., Ltd.) 2.0 Magnesium sulfate heptahydrate (FUJIFILM Wako Pure Chemical Corporation) 0.2 Manganese sulfate pentahydrate (Kishida Chemical Co., Ltd.) 0.05 Purified water Balance

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
EP22795395.7A 2021-04-28 2022-03-24 Wachstumsförderer für milchsäurekokken Pending EP4332213A1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
JP2021075882 2021-04-28
JP2021101500A JP7248250B2 (ja) 2021-04-28 2021-06-18 乳酸球菌の増殖促進剤
PCT/JP2022/013938 WO2022230476A1 (ja) 2021-04-28 2022-03-24 乳酸球菌の増殖促進剤

Publications (1)

Publication Number Publication Date
EP4332213A1 true EP4332213A1 (de) 2024-03-06

Family

ID=83847958

Family Applications (1)

Application Number Title Priority Date Filing Date
EP22795395.7A Pending EP4332213A1 (de) 2021-04-28 2022-03-24 Wachstumsförderer für milchsäurekokken

Country Status (3)

Country Link
US (1) US20240191177A1 (de)
EP (1) EP4332213A1 (de)
WO (1) WO2022230476A1 (de)

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63216475A (ja) * 1987-03-06 1988-09-08 Meiji Milk Prod Co Ltd 乳酸桿菌属細菌の自己溶解活性誘導法
NZ613335A (en) * 2010-12-28 2015-05-29 Kirin Holdings Kk Agent for inducing interferon production containing lactic acid bacteria
JP7506963B2 (ja) 2018-08-07 2024-06-27 株式会社ヤクルト本社 乳酸菌用培地
JP2020137456A (ja) 2019-02-28 2020-09-03 ビオフェルミン製薬株式会社 乳酸菌培養用培地

Also Published As

Publication number Publication date
WO2022230476A1 (ja) 2022-11-03
US20240191177A1 (en) 2024-06-13

Similar Documents

Publication Publication Date Title
Monika et al. Isolation and characterization of lactic acid bacteria from traditional pickles of Himachal Pradesh, India
Jung et al. Kimchi microflora: history, current status, and perspectives for industrial kimchi production
Witthuhn et al. Characterisation of the microbial population at different stages of Kefir production and Kefir grain mass cultivation
Mugula et al. Microbiological and fermentation characteristics of togwa, a Tanzanian fermented food
Kandler Carbohydrate metabolism in lactic acid bacteria
Williams et al. Energy sources of non-starter lactic acid bacteria isolated from Cheddar cheese
Van den Berghe et al. Enterocin A production by Enterococcus faecium FAIR-E 406 is characterised by a temperature-and pH-dependent switch-off mechanism when growth is limited due to nutrient depletion
Kaneko et al. Acetoin fermentation by citrate-positive Lactococcus lactis subsp. lactis 3022 grown aerobically in the presence of hemin or Cu2+
JP5940780B2 (ja) 石川県の伝統発酵食品から分離した乳酸菌、その培養物及びその利用
Díaz‐Muñiz et al. Lactobacillus casei metabolic potential to utilize citrate as an energy source in ripening cheese: a bioinformatics approach
Cardoso et al. Enhancement of trehalose production in dairy propionibacteria through manipulation of environmental conditions
Zamfir et al. Production kinetics of acidophilin 801, a bacteriocin produced by Lactobacillus acidophilus IBB 801
Lee et al. Microbial population dynamics and temperature changes during fermentation of kimjang kimchi
Liang et al. 3-Hydroxypropionic acid contributes to the antibacterial activity of glycerol metabolism by the food microbe Limosilactobacillus reuteri
Mei et al. Microbial diversity of a Camembert-type cheese using freeze-dried Tibetan kefir coculture as starter culture by culture-dependent and culture-independent methods
Skeie et al. Lactobacillus adjuncts in cheese: Their influence on the degradation of citrate and serine during ripening of a washed curd cheese
Ndagijimana et al. Growth and metabolites production by Penicillium brevicompactum in yoghurt
Chen et al. Evaluation of the spoilage heterogeneity of meat-borne Leuconostoc mesenteroides by metabonomics and in-situ analysis
Sugahara et al. Heterofermentative lactic acid bacteria such as Limosilactobacillus as a strong inhibitor of aldehyde compounds in plant-based milk alternatives
Yang et al. Substrate utilization by Clostridium estertheticum cultivated in meat juice medium
Shojaei Zinjanab et al. Natural flavor biosynthesis by lipase in fermented milk using in situ produced ethanol
Olaoye et al. Quantitative estimation of antimicrobials produced by lactic acid bacteria isolated from Nigerian beef.
Røssland et al. Influence of controlled lactic fermentation on growth and sporulation of Bacillus cereus in milk
Tomita et al. Characterisation of the bacterial community structures of sunki, a traditional unsalted pickle of fermented turnip leaves
Xiao et al. Isolation and characterization of plant-based lactic acid bacteria from spontaneously fermented foods using a new modified medium

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20231121

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)